Testosterone Parameter Assay Kit

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Multi-species Testosterone ELISA Standard Curve
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Product Details
Citations (50)
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Testosterone Parameter Assay Kit Summary

96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Serum (20 uL), EDTA Plasma (20 uL), Heparin Plasma (20 uL)
0.041 ng/mL
Assay Range
0.0 - 10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Measures Testosterone levels in various samples
Cross-reactivity observed with 1 or more available related molecules.Cross-species reactivity not tested.
Interference observed with 1 or more available related molecules.

Product Summary

The Parameter Testosterone kit is a 4.5 hour competitive enzyme immunoassay designed to measure Testosterone in cell culture supernates, serum, and plasma.


The recovery of Testosterone spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 103 95-117
EDTA Plasma (n=4) 94 85-107
Heparin Plasma (n=4) 92 84-101
Serum (n=4) 93 86-104


To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Testosterone were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Multi-species Testosterone ELISA Linearity

Scientific Data

Multi-species Testosterone ELISA Standard Curve

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Testosterone

Testosterone is an important secreted androgen synthesized from cholesterol-derived pregnenolone. Circulating testosterone is predominantly bound to proteins, including specific sex hormone binding globulin (SHBG) and nonspecific proteins such as albumin. It is believed that bioavailable testosterone includes the free steroid and the albumin-bound form. Testosterone is the main androgen secreted by the Leydig cells of the testes and affects both primary and secondary sexual development.

Alternate Names:
17b-Hydroxyandrost-4-ene-3-one; 17b-Testosterone ; Testosterone
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Primary Antibody Solution
  4.   Add 50 µL of Primary Antibody Solution to each well (except the non-specific binding (NSB) wells).
  5.   Incubate at room temperature for 1 hour on a horizontal orbital microplate shaker.
  6.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  7. Calibrator Diluent
  8.   Add 100 µL of Calibrator Diluent to the NSB wells.
  9.   Add 100 µL of Calibrator Diluent to the zero standard (B0) wells.

  10. 100 µL Standard, Control, or Sample
  11.   Add 100 µL of Standard, control, or sample to the remaining wells.

  12. 50 µL Conjugate
  13.   Add 50 µL of Conjugate to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours on the shaker.

  14. 200 µL Substrate Solution
  15. Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  16. 50 µL Stop Solution
  17. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Testosterone Parameter Assay Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

50 Citations: Showing 1 - 10
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  1. Small Molecule Cocktails Promote Fibroblast-to-Leydig-like Cell Conversion for Hypogonadism Therapy
    Authors: Yuan, F;Bai, K;Hou, Y;Zou, X;Sun, J;
    Species: Mouse, Human
    Sample Types: Cell Culture Supernates, Serum
  2. Eribulin mesylate induces bone mass loss by promoting osteoclastic bone resorption in mice
    Authors: Ishizaka, T;Horiuchi, K;Kondo, S;Isaji, M;Nakagawa, T;Inoue, M;Rikitake, H;Taguchi, E;Susa, M;Yoda, M;Ono, T;Kozai, Y;Chiba, K;
    Bone reports
    Species: Mouse
    Sample Types: Serum
  3. Effects of Ingesting Food Containing Heat-Killed Lactococcus lactis Strain Plasma on Fatigue and Immune-Related Indices after High Training Load: A Randomized, Double-Blind, Placebo-Controlled, and Parallel-Group Study
    Authors: Y Komano, K Fukao, K Shimada, H Naito, Y Ishihara, T Fujii, T Kokubo, H Daida
    Nutrients, 2023-04-04;15(7):.
    Species: Human
    Sample Types: Plasma
  4. Peroxiredoxin 5 regulates osteogenic differentiation through interaction with hnRNPK during bone regeneration
    Authors: E Cho, X Che, MJ Ang, S Cheon, J Lee, KS Kim, CH Lee, SY Lee, HY Yang, C Moon, C Park, JY Choi, TH Lee
    Elife, 2023-02-03;12(0):.
    Species: Mouse
    Sample Types: Serum
  5. Functional inhibition of deep brain non-visual opsins facilitates acute long day induction of reproductive recrudescence in male Japanese quail
    Authors: JH Pérez, E Tolla, VR Bishop, RG Foster, SN Peirson, IC Dunn, SL Meddle, TJ Stevenson
    Hormones and behavior, 2023-01-06;148(0):105298.
    Species: Quail
    Sample Types: Plasma
  6. Single-Cell RNAseq Resolve the Potential Effects of LanCL1 Gene in the Mouse Testis
    Authors: J Lu, J Liao, M Qin, H Li, Q Zhang, Y Chen, J Cheng
    Cells, 2022-12-19;11(24):.
    Species: Mouse
    Sample Types: Serum
  7. Agomelatine ameliorates cadmium-induced toxicity through the modification of HMGB-1/TLR-4/NFkappaB pathway
    Authors: HM Hafez, S Waz, NFG El-Tahawy, MZ Mohamed
    Toxicology and Applied Pharmacology, 2022-11-08;457(0):116313.
    Species: Rat
    Sample Types: Serum
  8. Upregulation of the oestrogen target gene SIX1 is associated with higher growth speed and decreased survival in HCV-positive women with hepatocellular carcinoma
    Authors: RM Critelli, F Milosa, A Romanzi, S Lasagni, G Marcelli, L Di Marco, A Pivetti, F Schepis, D Romagnoli, S Mancarella, F Dituri, ML Martinez-C, G Giannelli, E Villa
    Oncology Letters, 2022-09-21;24(5):395.
    Species: Human
    Sample Types: Serum
  9. Nestin-dependent mitochondria-ER contacts define stem Leydig cell differentiation to attenuate male reproductive ageing
    Authors: S Yao, X Wei, W Deng, B Wang, J Cai, Y Huang, X Lai, Y Qiu, Y Wang, Y Guan, J Wang
    Nature Communications, 2022-07-11;13(1):4020.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  10. Gene therapy with AR isoform 2 rescues spinal and bulbar muscular atrophy phenotype by modulating AR transcriptional activity
    Authors: WF Lim, M Forouhan, TC Roberts, J Dabney, R Elleringto, AA Speciale, R Manzano, M Lieto, G Sangha, S Banerjee, M Conceição, L Cravo, A Biscans, L Roux, N Pourshafie, C Grunseich, S Duguez, A Khvorova, M Pennuto, CJ Cortes, AR La Spada, KH Fischbeck, MJA Wood, C Rinaldi
    Science Advances, 2021-08-20;7(34):.
    Species: Mouse
    Sample Types: Serum
  11. The dynamic changes of Nrf2 mediated oxidative stress, DNA damage and base excision repair in testis of rats during aging
    Authors: H Zhao, L Song, N Ma, C Liu, Y Dun, Z Zhou, D Yuan, C Zhang
    Experimental gerontology, 2021-06-24;152(0):111460.
    Species: Rat
    Sample Types: Tissue Homogenates
  12. Endothelin-1 induces changes in the expression levels of steroidogenic enzymes and increases androgen receptor and testosterone production in the PC3 prostate cancer cell line
    Authors: MJ Torres, F López-Monc, D Herrera, S Indo, A Lefian, P Llanos, J Tapia, EA Castellón, HR Contreras
    Oncology reports, 2021-06-24;46(2):.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Melatonin Alleviated Potassium Dichromate-Induced Oxidative Stress and Reprotoxicity in Male Rats
    Authors: SAE Bashandy, H Ebaid, J Al-Tamimi, OA Ahmed-Fari, EA Omara, IM Alhazza
    BioMed Research International, 2021-06-15;2021(0):3565360.
    Species: Rat
    Sample Types: Plasma
  14. Adenine-induced chronic kidney disease induces a similar skeletal phenotype in male and female C57BL/6 mice with more severe deficits in cortical bone properties of male mice
    Authors: CE Metzger, EA Swallow, AJ Stacy, MR Allen
    PLoS ONE, 2021-04-23;16(4):e0250438.
    Species: Mouse
    Sample Types: Serum
  15. The androgen receptor/filamin A complex as a target in prostate cancer microenvironment
    Authors: M Di Donato, A Zamagni, G Galasso, E Di Zazzo, P Giovannell, MV Barone, M Zanoni, R Gunelli, M Costantini, F Auricchio, A Migliaccio, A Tesei, G Castoria
    Cell Death & Disease, 2021-01-26;12(1):127.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Microcystin-leucine arginine induced the apoptosis of GnRH neurons by activating the endoplasmic reticulum stress resulting in a decrease of serum testosterone level in mice
    Authors: H Jin, J Hou, X Meng, T Ma, B Wang, Z Liu, X Sha, J Ding, X Han
    Ecotoxicology and environmental safety, 2020-12-10;208(0):111748.
    Species: Mouse
    Sample Types: Serum
  17. Microbiota-dependent expansion of testicular IL-17-producing Vgamma6+ gammadelta T cells upon puberty promotes local tissue immune surveillance
    Authors: A Wilharm, HC Brigas, I Sandrock, M Ribeiro, T Amado, A Reinhardt, A Demera, L Hoenicke, T Strowig, T Carvalho, I Prinz, JC Ribot
    Mucosal Immunol, 2020-07-30;0(0):.
    Species: Mouse
    Sample Types: Serum
  18. Loss of p53 in mesenchymal stem cells promotes alteration of bone remodeling through negative regulation of osteoprotegerin
    Authors: T Velletri, Y Huang, Y Wang, Q Li, M Hu, N Xie, Q Yang, X Chen, Q Chen, P Shou, Y Gan, E Candi, AP Margherita, M Agostini, H Yang, G Melino, Y Shi, Y Wang
    Cell Death Differ., 2020-07-21;0(0):.
    Species: Mouse
    Sample Types: Serum
  19. Resistance Exercise in a Hot Environment Alters Serum Markers in Untrained Males
    Authors: A Eskandari, M Fashi, A Saeidi, D Boullosa, I Laher, A Ben Abderr, G Jabbour, H Zouhal
    Front Physiol, 2020-06-23;11(0):597.
    Species: Human
    Sample Types: Serum
  20. Icariin Improves Age-Related Testicular Dysfunction by Alleviating Sertoli Cell Injury via Upregulation of the ERalpha/Nrf2-Signaling Pathway
    Authors: H Zhao, X You, Q Chen, S Yang, Q Ma, Y He, C Liu, Y Dun, J Wu, C Zhang, D Yuan
    Front Pharmacol, 2020-05-12;11(0):677.
    Species: Rat
    Sample Types: Cell Lysates
  21. Targeted Deletion of PTEN in Kisspeptin Cells Results in Brain Region- and Sex-Specific Effects on Kisspeptin Expression and Gonadotropin Release
    Authors: AL Negrón, G Yu, U Boehm, M Acosta-Mar
    Int J Mol Sci, 2020-03-19;21(6):.
    Species: Mouse
    Sample Types: Plasma
  22. Development of Level A In Vitro-Vivo Correlation for Electrosprayed Microspheres Containing Leuprolide: Physicochemical, Pharmacokinetic, and Pharmacodynamic Evaluation
    Authors: DS Lee, DW Kang, GW Choi, HG Choi, HY Cho
    Pharmaceutics, 2020-01-02;12(1):.
    Species: Rat
    Sample Types: Plasma
  23. Impairment of learning and memory of mice offspring at puberty, young adulthood, and adulthood by low-dose Cd exposure during pregnancy and lactation via GABAAR ?5 and ? subunits
    Authors: Q Zhao, L Gao, Q Liu, Y Cao, Y He, A Hu, W Chen, J Cao, C Hu, L Li, F Tao
    Ecotoxicol. Environ. Saf., 2018-10-01;166(0):336-344.
    Species: Mouse
    Sample Types: Serum
  24. Testosterone represses urinary excretion of the alpha-tocopherol metabolite alpha-carboxymethylhydroxychroman in rats
    Authors: N Fujita, A Takenaka
    J. Nutr. Biochem., 2018-09-01;62(0):59-64.
    Species: Rat
    Sample Types: Plasma
  25. Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells
    Authors: Y Bai, C Zhu, M Feng, H Wei, L Li, X Tian, Z Zhao, S Liu, N Ma, X Zhang, R Shi, C Fu, Z Wu, S Zhang
    Stem Cell Res Ther, 2018-07-18;9(1):200.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  26. Serum cholesterol levels and tumor growth in a PTEN-null transgenic mouse model of prostate cancer
    Authors: EH Allott, EM Masko, AR Freedland, E Macias, K Pelton, KR Solomon, EA Mostaghel, GV Thomas, SV Pizzo, MR Freeman, SJ Freedland
    Prostate Cancer Prostatic Dis., 2018-05-23;0(0):.
    Species: Mouse
    Sample Types: Serum
  27. Pharmacokinetic-Pharmacodynamic Model for the Testosterone-Suppressive Effect of Leuprolide in Normal and Prostate Cancer Rats
    Authors: DS Lee, SJ Kim, GW Choi, YB Lee, HY Cho
    Molecules, 2018-04-15;23(4):.
    Species: Rat
    Sample Types: Plasma
  28. Effect of speed endurance training and reduced training volume on running economy and single muscle fiber adaptations in trained runners
    Authors: C Skovgaard, D Christians, PM Christense, NW Almquist, M Thomassen, J Bangsbo
    Physiol Rep, 2018-02-01;6(3):.
    Species: Human
    Sample Types: Plasma
  29. Reproductive Hormones and Their Receptors May Affect Lung Cancer
    Authors: M Dou, K Zhu, Z Fan, Y Zhang, X Chen, X Zhou, X Ding, L Li, Z Gu, M Guo, M Yan, X Deng, P Shen, S Wang
    Cell. Physiol. Biochem., 2017-11-30;44(4):1425-1434.
    Species: Mouse
    Sample Types: Serum
  30. Testosterone in advance age: a New Zealand longitudinal cohort study: Life and Living in Advanced Age (Te Pu?waitanga o Ng? Tapuwae Kia Ora Tonu)
    Authors: MJ Connolly, N Kerse, T Wilkinson, O Menzies, A Rolleston, YH Chong, JB Broad, SA Moyes, S Jatrana, R Teh
    BMJ Open, 2017-11-12;7(11):e016572.
    Species: Human
    Sample Types: Serum
  31. Lysophosphatidic acid acyltransferase 3 tunes the membrane status of germ cells by incorporating docosahexaenoic acid during spermatogenesis
    Authors: Y Iizuka-His, D Hishikawa, J Sasaki, K Takubo, M Goto, K Nagata, H Nakanishi, H Shindou, T Okamura, C Ito, K Toshimori, T Sasaki, T Shimizu
    J. Biol. Chem., 2017-06-03;0(0):.
    Species: Mouse
    Sample Types: Serum
  32. Phospholipase C-related but Catalytically Inactive Proteins Regulate Ovarian Follicle Development
    Authors: M Matsuda, M Hirata
    J. Biol. Chem., 2017-03-30;0(0):.
    Species: Mouse
    Sample Types: Serum
  33. Transplantation of CD51(+) stem Leydig cells: A new strategy for the treatment of testosterone deficiency
    Authors: ZJ Zang, J Wang, Z Chen, Y Zhang, Y Gao, Z Su, Y Tuo, Y Liao, M Zhang, Q Yuan, C Deng, MH Jiang, AP Xiang
    Stem Cells, 2017-03-05;0(0):.
    Species: Mouse
    Sample Types: Serum
  34. Effects of Microcystis on Hypothalamic-Pituitary-Gonadal-Liver Axis in Nile Tilapia (Oreochromis niloticus)
    Authors: J Chen, S Meng, H Xu, Z Zhang, X Wu
    Bull Environ Contam Toxicol, 2017-02-24;0(0):.
    Species: Fish - Oreochrmois niloticus (Tilapia)
    Sample Types: Serum
  35. Effect of lipid metabolism on male fertility
    Authors: N Kim, H Nakamura, H Masaki, K Kumasawa, KI Hirano, T Kimura
    Biochem. Biophys. Res. Commun, 2017-02-22;0(0):.
    Species: Human
    Sample Types: Plasma
  36. Effect of increased and maintained frequency of speed endurance training on performance and muscle adaptations in runners
    Authors: Jens Bangsbo
    J. Appl. Physiol., 2016-11-17;0(0):jap.00537.201.
    Species: Human
    Sample Types: Plasma
  37. Cutting edge: c-Kit signaling differentially regulates type 2 innate lymphoid cell accumulation and susceptibility to central nervous system demyelination in male and female SJL mice.
    Authors: Russi A, Walker-Caulfield M, Ebel M, Brown M
    J Immunol, 2015-05-13;194(12):5609-13.
    Species: Mouse
    Sample Types: Serum
  38. Diet-induced obesity in male C57BL/6 mice decreases fertility as a consequence of disrupted blood-testis barrier.
    Authors: Fan, Yong, Liu, Yue, Xue, Ke, Gu, Guobao, Fan, Weimin, Xu, Yali, Ding, Zhide
    PLoS ONE, 2015-04-17;10(4):e0120775.
    Species: Mouse
    Sample Types: Serum
  39. Alterations of bone density, microstructure, and strength of the distal radius in male patients with rheumatoid arthritis: a case-control study with HR-pQCT.
    Authors: Zhu T, Griffith J, Qin L, Hung V, Fong T, Au S, Li M, Lam Y, Wong C, Kwok A, Leung P, Li E, Tam L
    J Bone Miner Res, 2014-09-01;29(9):2118-29.
    Species: Human
    Sample Types: Serum
  40. Inhibitor of differentiation 4 (ID4) inactivation promotes de novo steroidogenesis and castration-resistant prostate cancer.
    Authors: Patel, Divya, Knowell, Ashley E, Korang-Yeboah, Maxwell, Sharma, Pankaj, Joshi, Jugal, Glymph, Shanora, Chinaranagari, Swathi, Nagappan, Peri, Palaniappan, Ravi, Bowen, Nathan J, Chaudhary, Jaideep
    Mol Endocrinol, 2014-06-12;28(8):1239-53.
    Species: Human
    Sample Types: Cell Culture Supernates
  41. Osteopontin deletion prevents the development of obesity and hepatic steatosis via impaired adipose tissue matrix remodeling and reduced inflammation and fibrosis in adipose tissue and liver in mice.
    Authors: Lancha A, Rodriguez A, Catalan V, Becerril S, Sainz N, Ramirez B, Burrell M, Salvador J, Fruhbeck G, Gomez-Ambrosi J
    PLoS ONE, 2014-05-28;9(5):e98398.
    Species: Mouse
    Sample Types: Serum
  42. Antagonistic effects of a mixture of low-dose nonylphenol and di-n-butyl phthalate (monobutyl phthalate) on the Sertoli cells and serum reproductive hormones in prepubertal male rats in vitro and in vivo.
    Authors: Hu, Yang, Wang, Ruoyu, Xiang, Zou, Qian, Weiping, Han, Xiaodong, Li, Dongmei
    PLoS ONE, 2014-03-27;9(3):e93425.
    Species: Rat
    Sample Types: Serum
  43. Insulin-like 3-induced rat preantral follicular growth is mediated by growth differentiation factor 9.
    Authors: Xue, Kai, Kim, Ji Young, Liu, Jia-yin, Tsang, Benjamin
    Endocrinology, 2013-12-20;155(1):156-67.
    Species: Rat
    Sample Types: Cell Culture Supernates
  44. Host-derived adiponectin is tumor-suppressive and a novel therapeutic target for multiple myeloma and the associated bone disease.
    Authors: Fowler JA, Lwin ST, Drake MT
    Blood, 2011-09-08;118(22):5872-82.
    Species: Human
    Sample Types: Serum
  45. Endocrine responses during overnight recovery from exercise: impact of nutrition and relationships with muscle protein synthesis.
    Authors: Betts JA, Beelen M, Stokes KA, Saris W HM, van Loon L JC
    Int J Sport Nutr Exerc Metab, 2011-08-10;21(5):398-409.
    Species: Human
    Sample Types: Plasma
  46. Inhibitor of DNA binding 4 is expressed selectively by single spermatogonia in the male germline and regulates the self-renewal of spermatogonial stem cells in mice.
    Authors: Oatley MJ, Kaucher AV, Racicot KE
    Biol. Reprod., 2011-05-04;85(2):347-56.
    Species: Mouse
    Sample Types: Serum
  47. Testosterone regulates cell proliferation in aggressive fibromatosis (desmoid tumour).
    Authors: Hong H, Nadesan P, Poon R, Alman BA
    Br. J. Cancer, 2011-04-05;104(9):1452-8.
    Species: Mouse
    Sample Types: Serum
  48. Skeletal muscle myofibrillar and sarcoplasmic protein synthesis rates are affected differently by altitude-induced hypoxia in native lowlanders.
    Authors: Holm L, Haslund ML, Robach P, van Hall G, Calbet JA, Saltin B, Lundby C
    PLoS ONE, 2010-12-20;5(12):e15606.
    Species: Human
    Sample Types: Plasma
  49. Castration Differentially Alters Basal and Leucine-Stimulated Tissue Protein Synthesis in Skeletal Muscle and Adipose Tissue.
    Authors: Jiao Q, Pruznak AM, Huber D, Vary TC, Lang CH
    Am. J. Physiol. Endocrinol. Metab., 2009-09-15;0(0):.
    Species: Rat
    Sample Types: Plasma
  50. Inhibition of tumor growth progression by antiandrogens and mTOR inhibitor in a Pten-deficient mouse model of prostate cancer.
    Authors: Zhang W, Zhu J, Efferson CL, Ware C, Tammam J, Angagaw M, Laskey J, Bettano KA, Kasibhatla S, Reilly JF, Sur C, Majumder PK
    Cancer Res., 2009-09-08;69(18):7466-72.
    Species: Mouse
    Sample Types: Plasma


  1. Does R&D Systems® Testosterone Parameter Assay Kit, Catalog # KGE010, detect free or total testosterone?

    • 'Free' testosterone is typically defined as testosterone which is bioavailable, and not bound to sex hormone binding globulin (SHBG). Interference testing performed by spiking SHBG into testosterone-containing samples, demonstrates interference in testosterone detection with Catalog # KGE010, when the concentration of SHBG is greater than 100 ng/ml. (For further details on interference testing, please refer to the "Specificity" section in the product insert.) 

      Assays for 'total' testosterone measure both 'free' testostrone as well as testosterone bound to SHBG. Since KGE010 does not detect testosterone bound to SHBG, this assay does not measure 'total' testosterone.

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Testosterone Parameter Assay Kit
By Anonymous on 03/22/2018
Sample Tested: Serum,Serum-free Cell Culture Media